Prevalence, risk factors and zoonotic potential of intestinal parasites in dogs from four locations in Morocco.

Dogs can harbor various intestinal parasites that have serious clinical, economic, and zoonotic impact. In Morocco, the epidemiological status of those parasites is largely unknown. This study aimed to obtain data on the prevalence of intestinal parasites in various Moroccan dog populations, to identify associated risk factors, to evaluate people's knowledge regarding zoonotic parasites and to estimate the risk of human infection. A total of 291 fecal samples were analyzed using a 33% Zinc Sulphate (ZnSO4) centrifugal flotation and a sheather's sugar simple flotation techniques. In addition, 100 dog owners were asked to fill out a questionnaire about their knowledge regarding canine zoonotic intestinal parasites. Overall, 58% of sampled dogs were positive for at least one parasite species. Ancylostoma/Uncinaria spp. (31.9%), Toxascaris leonina (27.4%), Toxocara canis (27.1%), Cystoisospora spp. (13.4%) and Giardia spp. (7.2%) were the most frequently isolated parasites. The overall prevalence was associated with the dogs' activity, and feces consistency. More specifically, the prevalence of Ancylostoma/Uncinaria spp., Toxascaris leonina and Cystoisospora spp. was associated with dogs' activity and feces consistency and, additionally with age for Cystoisosporaspp.Toxocara canis prevalence was associated with age and feces consistency, while Giardia spp. prevalence was associated with dogs' activity. A weak awareness among dog owners regarding the zoonotic potential of canine intestinal parasites was noticed (33%) especially when their knowledge was compared to another endemic zoonotic disease, i.e. rabies (85%). Furthermore, the rate of respecting the vaccination protocol (82%) was significantly higher than the rate of respecting the deworming protocol (47%). We conclude that intestinal parasites, including the zoonotic ones, were highly prevalent in the dogs sampled in this study whereas people's knowledge about those parasites was very limited. These findings suggest that the risk of human infection is very likely, which highlights the need for effective control programs and health education.

Canine Leishmaniasis in Morocco: A Descriptive Prospective Clinical Study.

Canine leishmaniasis (CanL) is a zoonotic vector-borne disease that is endemic in the Mediterranean Basin including Morocco. Dogs play a major epidemiological role in this zoonosis as reservoir hosts. This study investigated the clinical manifestations of CanL in dogs naturally infected with Leishmania infantum. A total of 96 dogs presented to the Small Animal Clinic of the Hassan II Agronomy and Veterinary Institute (IAV Hassan II) of Rabat, Morocco, and were tested by RT-PCR and/or serology. Among them, 32 (33.3%) were positive to Leishmania infantum infection. The majority of the positive dogs (93.7%) came from urban areas. Most of them were male (62.5%) and purebreds (65.6%), were aged between 3 and 7 years (71.8%), and had outside activities (guarding, hunting, livestock guarding, and service activities) (71.8%) and all of them were living exclusively outdoor or had free access to the outdoor environment. Lymphadenomegaly (81.2%), dermatological disorders (65.6%) (mostly exfoliative dermatitis), weight loss (59.3%), exercise intolerance (56.2%), anorexia (28.1%), hyporexia (15.6%), and ocular lesions (28.1%) were the most frequent clinical signs and complaints recorded. Anemia and hyperproteinemia due to hyperglobulinemia were observed in 68.7% and 72.7% of the cases, respectively. These results suggest that CanL leads to various nonspecific clinical signs as described previously, making the diagnosis challenging. Since CanL is endemic in Morocco, it should be recommended to systematically test dogs displaying clinical signs compatible with this disease and to regularly screen asymptomatic at-risk dogs. It is also crucial to educate dog owners about the zoonotic aspect of the disease and to encourage intersectorial collaboration following the "One Health" concept, in order to contribute to a more effective control/prevention of human and canine leishmaniasis.

Epidemiological and Clinicopathological Features of Anaplasma phagocytophilum Infection in Dogs: A Systematic Review.

Anaplasma phagocytophilum is a worldwide emerging zoonotic tick-borne pathogen transmitted by Ixodid ticks and naturally maintained in complex and incompletely assessed enzootic cycles. Several studies have demonstrated an extensive genetic variability with variable host tropisms and pathogenicity. However, the relationship between genetic diversity and modified pathogenicity is not yet understood. Because of their proximity to humans, dogs are potential sentinels for the transmission of vector-borne pathogens. Furthermore, the strong molecular similarity between human and canine isolates of A. phagocytophilum in Europe and the USA and the positive association in the distribution of human and canine cases in the USA emphasizes the epidemiological role of dogs. Anaplasma phagocytophilum infects and survives within neutrophils by disregulating neutrophil functions and evading specific immune responses. Moreover, the complex interaction between the bacterium and the infected host immune system contribute to induce inflammatory injuries. Canine granulocytic anaplasmosis is an acute febrile illness characterized by lethargy, inappetence, weight loss and musculoskeletal pain. Hematological and biochemistry profile modifications associated with this disease are unspecific and include thrombocytopenia, anemia, morulae within neutrophils and increased liver enzymes activity. Coinfections with other tick-borne pathogens (TBPs) may occur, especially with Borrelia burgdorferi, complicating the clinical presentation, diagnosis and response to treatment. Although clinical studies have been published in dogs, it remains unclear if several clinical signs and clinicopathological abnormalities can be related to this infection.

Molecular Characterization of Tandem Repeat Protein 36 Gene of Ehrlichia canis Detected in Naturally Infected Dogs from Peru.

Ehrlichia spp. are emerging infectious pathogens, especially in the Americas. Although Ehrlichia canis is primarily a parasite of dogs, polymerase chain reaction-confirmed human infections have been reported from Mexico, Venezuela, and Costa Rica. This study reports the presence of E. canis DNA in 13.7% of 205 dogs from urban areas in Peru and of those, five were analyzed for phylogenetic variation using the Tandem Repeat Protein 36 (TRP36) gene. The use of the TRP36 gene for such analysis was validated against 16S rRNA and heat shock protein genes using Shannon's entropy bioinformatic approach. When compared with other E. canis strains previously reported, three unique and novel E. canis strains were detected. In addition, the TRP36 amino acid tandem repeat sequences of the Peruvian strains share close similarity to an E. canis strain detected from four human blood bank samples in Costa Rica. This study reports for the first time domestic dogs infected with E. canis strains closely related to a zoonotic strain, which may be of public health concern as dogs can be chronically infected with this pathogen.

Anaplasma spp. in dogs and owners in north-western Morocco.

BACKGROUND: Anaplasma phagocytophilum is an emerging tick-borne zoonotic pathogen of increased interest worldwide which has been detected in northern Africa. Anaplasma platys is also present in this region and could possibly have a zoonotic potential. However, only one recent article reports on the human esposure to A. phagocytophilum in Morocco and no data are available on canine exposure to both bacteria. Therefore, we conducted a cross-sectional epidemiological study aiming to assess both canine and human exposure to Anaplasma spp. in Morocco. A total of 425 dogs (95 urban, 160 rural and 175 working dogs) and 11 dog owners were sampled from four cities of Morocco. Canine blood samples were screened for Anaplasma spp. antibodies by an enzyme-linked immunosorbent assay (ELISA) and for A. phagocytophilum and A. platys DNA by a real-time polymerase chain reaction (RT-PCR) targeting the msp2 gene. Human sera were tested for specific A. phagocytophilum immunoglobulin G (IgG) using a commercial immunofluorescence assay (IFA) kit. RESULTS: Anaplasma spp. antibodies and A. platys DNA were detected in 21.9 and 7.5% of the dogs, respectively. Anaplasma phagocytophilum DNA was not amplified. Anaplasma platys DNA was significantly more frequently amplified for working dogs. No statistically significant differences in the prevalence of Anaplasma spp. antibodies or A. platys DNA detection were observed between sexes, age classes or in relation to exposure to ticks. A total of 348 Rhipicephalus sanguineus (sensu lato) ticks were removed from 35 urban and working dogs. The majority of dog owners (7/10) were seroreactive to A. phagoyctophilum IgG (one sample was excluded because of hemolysis). CONCLUSIONS: This study demonstrates the occurrence of Anaplasma spp. exposure and A. platys infection in dogs, and A. phagocytophilum exposure in humans in Morocco.

Detection of Anaplasma spp. and Ehrlichia spp. anibodies, and Dirofilaria immitis antigens in dogs from seven locations of Morocco.

In Morocco no data has been published on canine exposure to Anaplasma spp., Borrrelia burgdorferi, and Ehrlichia spp., and only one report is available on the occurrence of Dirofilaria immitis in dogs. Therefore, the aim of this study was to collect current data on the canine exposure to these vector-borne pathogens (VBPs) in Morocco. A total of 217 urban (n=57), rural (n=110) and military (n=50) dogs from seven Moroccan locations were screened for Anaplasma spp., B. burgdorferi and Ehrlichia spp. antibodies and for D. immitis antigens using a commercial in-clinic ELISA test. Of these dogs, 182 (83.9%) tested positive for at least one pathogen and positivity to two or three pathogens was found in 14.3% and 2.3% of the dogs, respectively. Ehrlichia spp. antibodies (34.6%) were the most frequently detected followed by Anaplasma spp. antibodies (16.6%) and D. immitis antigens (16.1%). None of the dogs was tested seropositive to B. burgdorferi. Statistically significant differences in seropositivity rates were found for Ehrlichia spp. and D. immitis in rural dogs especially those from the north central region (p<0.001) but not for Anaplasma spp. No significant difference was found according to the health status of the dog. This study demonstrates that Moroccan dogs are at high risk of acquiring a vector-borne infection.

Human Exposure to Anaplasma phagocytophilum in Two Cities of Northwestern Morocco.

Anaplasma phagocytophilum is an emerging tick-borne zoonosis with extensive increased interest. Epidemiological data are available in several regions of the USA, Europe and Asia in contrast to other parts of the world such as North Africa. Blood samples of 261 healthy individuals divided in two groups i.e., dog handlers and blood donors were analysed. Indirect immunofluorescent assay using a commercial kit was performed to detect specific A. phagocytophilum IgG. Two dilutions were used to assess the prevalence of seroreactive samples. Demographic variables were assessed as potential risk factors using exact logistic regression. Seropositivity rates reached 37% and 27% in dog handlers and 36% and 22% in blood donors. No statistically significant differences were found in the prevalence rates between the two groups. Analysis of risk factors such as gender, age groups, outdoor activities, self-reported previous exposure to ticks, or contact with domestic animals (dogs, cats, ruminants and horses) did not shown any significant difference. A. phagocytophilum exposure was common in both high-risk population and blood donors in Morocco.

Modulatory influences of estradiol and other anorexigenic hormones on metabotropic, Gi/o-coupled receptor function in the hypothalamic control of energy homeostasis.

The appetite suppressant actions of estradiol are due to its ability to attenuate orexigenic signals and potentiate anorexigenic signals. The work from my laboratory has shown that male guinea pigs are more sensitive to the hyperphagic and hypothermic effects of cannabinoids than their female counterparts. Cannabinoid sensitivity is further dampened by the activational effects of estradiol. This occurs via the hypothalamic feeding circuitry, where estradiol rapidly attenuates the cannabinoid CB1 receptor-mediated presynaptic inhibition of glutamatergic input onto anorexigenic proopiomelanocortin (POMC) neurons in the arcuate nucleus. This disruption is blocked by the estrogen receptor antagonist ICI 182,780, and associated with increased expression of phosphatidylinositol-3-kinase (PI3K). Moreover, the ability of estradiol to reduce both the cannabinoid-induced hyperphagia and glutamate release onto POMC neurons is abrogated by the PI3K inhibitor PI 828. The peptide orphanin FQ/nociceptin (OFQ/N) activates opioid receptor-like (ORL)1 receptors to hyperpolarize and inhibit POMC neurons via the activation of postsynaptic G protein-gated, inwardly-rectifying (GIRK) channels. We have demonstrated that the fasting-induced hyperphagia observed in ORL1-null mice is blunted compared to wild type controls. In addition, the ORL1 receptor-mediated activation of GIRK channels in POMC neurons from ovariectomized female rats is markedly impaired by estradiol. The estrogenic attenuation of presynaptic CB1 and postsynaptic ORL1 receptor function may be part of a more generalized mechanism through which anorexigenic hormones suppress orexigenic signaling. Indeed, we have found that leptin robustly suppresses the OFQ/N-induced activation of GIRK channels in POMC neurons. Furthermore, its ability to augment excitatory input onto POMC neurons is blocked by PI 828. Thus, estradiol and other hormones like leptin reduce energy intake at least partly by activating PI3K to disrupt the pleiotropic functions of Gi/o-coupled receptors that inhibit anorexigenic POMC neurons.

Dog population management for the control of human echinococcosis.

Cystic and alveolar hydatid disease of humans caused by infection with Echinococcus granulosus or Echinococcus multilocularis are significant zoonoses in developing countries. For human infections, the main definitive host is the dog, and reduction in the population of unwanted dogs, together with anthelmintic treatment of wanted dogs, are recommended control procedures for these zoonoses. Both owned and unowned dogs have been shown to be a major source of Echinococcus spp. infection in developing countries. Unowned dogs are the most challenging category in dog population management for the control of major zoonotic diseases. Unowned dogs are those dogs that do not have an owner, and those dogs whose owner cannot readily be identified. Control of numbers of unowned dogs can be done in various ways if funds are available. Fertility control and humane euthanasia are likely to be the most effective procedures in developing countries. Fertility control requires significant funding, and where resources are scarce humane euthanasia may be the most effective option. Both procedures are ongoing events, with no predictable end point. This paper examines the sociology and technology for the population management of owned and unowned dogs, specifically for the reduction of human hydatid disease. Examples are given for developing and developed countries. Although a "One Health" approach is desirable, the technology for hydatid control is different from that for rabies, and FAO Animal Welfare recommendations for dog population management should be adjusted accordingly.

Toward proof of concept of a one health approach to disease prediction and control.

A One Health approach considers the role of changing environments with regard to infectious and chronic disease risks affecting humans and nonhuman animals. Recent disease emergence events have lent support to a One Health approach. In 2010, the Stone Mountain Working Group on One Health Proof of Concept assembled and evaluated the evidence regarding proof of concept of the One Health approach to disease prediction and control. Aspects examined included the feasibility of integrating human, animal, and environmental health and whether such integration could improve disease prediction and control efforts. They found evidence to support each of these concepts but also identified the need for greater incorporation of environmental and ecosystem factors into disease assessments and interventions. The findings of the Working Group argue for larger controlled studies to evaluate the comparative effectiveness of the One Health approach.

Infection of domestic dogs in peru by zoonotic bartonella species: a cross-sectional prevalence study of 219 asymptomatic dogs.

Bartonella species are emerging infectious organisms transmitted by arthropods capable of causing long-lasting infection in mammalian hosts. Among over 30 species described from four continents to date, 15 are known to infect humans, with eight of these capable of infecting dogs as well. B. bacilliformis is the only species described infecting humans in Peru; however, several other Bartonella species were detected in small mammals, bats, ticks, and fleas in that country. The objective of this study was to determine the serological and/or molecular prevalence of Bartonella species in asymptomatic dogs in Peru in order to indirectly evaluate the potential for human exposure to zoonotic Bartonella species. A convenient sample of 219 healthy dogs was obtained from five cities and three villages in Peru. EDTA-blood samples were collected from 205 dogs, whereas serum samples were available from 108 dogs. The EDTA-blood samples were screened by PCR followed by nucleotide sequencing for species identification. Antibodies against B. vinsonii berkhoffii and B. rochalimae were detected by IFA (cut-off of 1∶64). Bartonella DNA was detected in 21 of the 205 dogs (10%). Fifteen dogs were infected with B. rochalimae, while six dogs were infected with B. v. berkhoffii genotype III. Seropositivity for B. rochalimae was detected in 67 dogs (62%), and for B. v. berkhoffii in 43 (40%) of the 108 dogs. Reciprocal titers ≥1∶256 for B. rochalimae were detected in 19% of dogs, and for B. v. berkhoffii in 6.5% of dogs. This study identifies for the first time a population of dogs exposed to or infected with zoonotic Bartonella species, suggesting that domestic dogs may be the natural reservoir of these zoonotic organisms. Since dogs are epidemiological sentinels, Peruvian humans may be exposed to infections with B. rochalimae or B. v. berkhoffii.

Estradiol negatively modulates the pleiotropic actions of orphanin FQ/nociceptin at proopiomelanocortin synapses.

Orphanin FQ/nociceptin (OFQ/N) inhibits the activity of proopiomelanocortin (POMC) neurons located in the hypothalamic arcuate nucleus (ARH) that regulate female sexual behavior and energy balance. We tested the hypothesis that estradiol modulates the ability of OFQ/N to pre- and postsynaptically decrease the excitability of these cells. To this end, whole-cell patch-clamp recordings were performed in hypothalamic slices prepared from ovariectomized rats, including some that were injected with the retrograde tracer Fluorogold in the medial preoptic nucleus (MPN) to label the POMC neurons regulating sexual receptivity. OFQ/N (1 µM) evoked a robust outward current in ARH neurons from vehicle-treated animals that was blocked by the opioid receptor-like (ORL)1 receptor antagonist UFP-101 (100 nM) and the G protein-gated, inwardly rectifying K⁺ (GIRK-1) channel blocker tertiapin (10 nM). OFQ/N also produced a decrease in the frequency of glutamatergic, miniature excitatory postsynaptic currents (mEPSCs), which was also antagonized by UFP-101. Estradiol benzoate (2 µg) increased basal mEPSC frequency and markedly diminished both the OFQ/N-induced activation of postsynaptic GIRK-1 channel currents and the presynaptic inhibition of glutamatergic neurotransmission. These effects were observed in identified POMC neurons, including eight that projected to the MPN. Taken together, these data reveal that estradiol attenuates the pleiotropic inhibitory actions of OFQ/N on POMC neurons: presynaptically through reducing the OFQ/N inhibition of glutamate release and postsynaptically by reducing ORL1 signaling through GIRK channels. As such, they impart critical insight into a mechanism for estradiol to increase the activity of POMC neurons that inhibit sexual receptivity.

Factores domiciliarios asociados con la presencia de hidatidosis humana en tres comunidades rurales de Junín, Perú / Household factors associated with the presence of human hydatid disease in three rural communities of Junin, Peru

Introducción. La hidatidosis, zoonosis producida por el estadio larvario de la taenia Echinococcus granulosus, es un problema de salud pública a nivel nacional, en especial en aquellas regiones dedicadas a la crianza de ganado. A la fecha, se han descrito factores, a nivel individual, asociados con la infección por E. granulosus; sin embargo, no se encontró reporte previo que explore la asociación entre características de la vivienda y la presencia de esta enfermedad en alguno de sus miembros. Objetivos. Explorar la asociación entre las características de la vivienda y la presencia de hidatidosis entre las personas que la habitan. Materiales y métodos. Se realizó un estudio transversal en viviendas de tres comunidades rurales de Junín. Se evaluó el grado de asociación entre las características de la vivienda y la presencia de hidatidosis en el hogar mediante un análisis de regresión logística múltiple (RLM). Resultados. De un total de 417 viviendas evaluadas, 56 (13 por ciento) de ellas tenían al menos un caso positivo entre sus miembros; luego del análisis de RLM se observó que aquellas viviendas con más de tres miembros, localizadas en la comunidad con quintil de pobreza más bajo, que refirieron crianza de animales, y con una cobertura de evaluación mayor al 25 por ciento presentaron una mayor probabilidad de tener al menos un caso de positivo entre sus miembros. Conclusión. Las características observadas deben ser tomadas en cuenta para la determinación preliminar de subgrupos de alto riesgo, optimizando así el uso de los recursos y mejorando la eficacia de los programas de despistaje.

Introduction. Hydatid disease, a zoonosis caused by the larval stage of the parasite Echinococcus granulosus, is a public health problem at national level, especially in those regions dedicated to raising livestock. By now, there are many factors, at individual level, that have been associated to the infection by E. granulosus; nevertheless there is not any previous report that explore the association between household characteristics and the presence of the disease among household members. Objective. To explore the association between household characteristics and the presence of hydatid disease among household members. Material and methods. We performed a cross-sectional study in the households of 3 rural communities located in Junín, we evaluated the association between household characteristics and the presence of hydatid disease by multiple logistic regression (MLR). Results. From 417 evaluated households, 56 (13 percent) of them had at least one positive case among its members. Multivariate analysis showed that households with three or more members, located in the community with the lowest quintile of poverty, that reported raising livestock, and with evaluation coverage greater than 25 percent were more likely to have at least one positive case among its members. Conclusion. The observed characteristics be taken into account in the preliminary definition of high-risk subgroups, optimizing the use of resources and improving the effectiveness of screening programs.

[Household factors associated with the presence of human hydatid disease in three rural communities of Junin, Peru]. / Factores domiciliarios asociados con la presencia de hidatidosis humana en tres comunidades rurales de Junín, Perú.

INTRODUCTION: Hydatid disease, a zoonosis caused by the larval stage of the parasite Echinococcus granulosus, is a public health problem at national level, especially in those regions dedicated to raising livestock. By now, there are many factors, at individual level, that have been associated to the infection by E. granulosus; nevertheless there is not any previous report that explore the association between household characteristics and the presence of the disease among household members. OBJECTIVE: To explore the association between household characteristics and the presence of hydatid disease among household members. MATERIAL AND METHODS: We performed a cross-sectional study in the households of 3 rural communities located in Junín, we evaluated the association between household characteristics and the presence of hydatid disease by multiple logistic regression (MLR). RESULTS: From 417 evaluated households, 56 (13%) of them had at least one positive case among its members. Multivariate analysis showed that households with three or more members, located in the community with the lowest quintile of poverty, that reported raising livestock, and with evaluation coverage greater than 25% were more likely to have at least one positive case among its members. CONCLUSION: The observed characteristics be taken into account in the preliminary definition of high-risk subgroups, optimizing the use of resources and improving the effectiveness of screening programs.

Antibodies to Bartonella vinsonii subsp. Berkhoffii in Moroccan dogs.

Bartonella vinsonii subsp. berkhoffii is a fastidious microorganism that has been recognized as an emerging human and canine pathogen. We report for the first time on the prevalence of antibodies to B. vinsonii subsp. berkhoffii in domestic dogs from Morocco. The overall seroprevalence was 38% (56 of 147 dogs tested). Most of the seropositive dogs were stray dogs from Rabat (36%, 8 of 22) and Khenifra (47%, 47 of 101). Antibodies against B. vinsonii subsp. berkhoffii were found infrequently among pet dogs from Rabat (4%, 1 of 24).